Microsoft office excel 2016

Normality of the datasets obtained from each experiment was tested using the Shapiro–Wilk test; however, if equal variance failed, Friedman nonparametric and pairwise post hoc tests (Bonferroni) were used. Indeed, as equal variance failed in IFE, we analyzed it as a nonparametric dataset. One-way analysis of variance with Bonferroni’s post hoc test was used to compare the time course changes in VO₂, VCO₂, RER, and GE. For a graphical representation of these data, we represented the mean and individual values as solid and dotted lines, respectively. Microsoft Office Excel 2016 and Prism v. 9.0 (GraphPad Inc., La Jolla, CA, USA) were used for data processing and statistical analyses, respectively. Statistical significance was set at p < 0.05.

Multidimensional scaling (MDS) analysis was performed using all the protein-coding genes to cluster the samples according to the overall similarity of their gene expression patterns to determine whether the gene expression patterns of the phenotypic classes could be clearly distinguished. For MDS analysis, the pairwise distances between the samples were determined using the function “dist” (maximum distance measure) and were plotted using R. The log_2-transformed values were used for this analysis, and rows with zero expression in all samples were eliminated. Hierarchical clustering was performed, and a heatmap was generated using “heatmap.2” (ward.D2 clustering function) in the gplots package (v3.1.1) to determine the phenotypic grouping and the specific gene expression patterns. All experiments were performed in triplicate. Data are presented as the mean ± SD values. Data were processed in Microsoft Office Excel 2016 and GraphPad Prism software version 8.1.1 (Prism, La Jolla, CA, USA). Statistical analyses were performed using t tests. Statistical significance was set at P ≤ 0.05.

Sample size was chosen based on the need for statistical power. All experiments were performed triplicate. Data represent mean ± S.D. Data were processed in Microsoft Office Excel 2016 and GraphPad Prism software version 8.1.1 (Prism, La Jolla, CA). Statistical analysis was performed using a t-test. P ≤ 0.05 was considered statistically significant.